Tumor necrosis factor-alpha (TNF alpha) is thought to play a physiological role in the brain. These studies were performed to determine whether a diurnal rhythm of TNF alpha exist in the rat brain. Samples were collected from hippocampus, hypothalamus, cerebral cortex, cerebellum, pons and midbrain at light onset and at 6 h intervals thereafter over a day. A TNF alpha bioassay was used to measure TNF alpha in each area. TNF alpha was highest at light onset in the hypothalamus, hippocampus and cerebral cortex. Levels at light onset were about 10-fold greater than minimal night-time levels. Changes in TNF alpha activity in other brain areas were also evident, but smaller. These results support the hypothesis that TNF alpha has physiological roles in the brain.